0.5 ml PCR tubes (Only when your PCR thermocycler cannot accommodate 0.2 ml tubes/strips) Cut segments of non-skirted 96-well plates

Transcription

1 Submission of Samples under Form B (i.e. to be sequenced or cleaned up by us) Fewer than 16 Submit samples in one of the following: 0.2 ml PCR tubes 0.2 ml high/low profile 8/12-tube strips Cut segments of non-skirted 96-well plates 0.5 ml PCR tubes (Only when your PCR thermocycler cannot accommodate 0.2 ml tubes/strips) More than 16 but fewer than 30 Submit samples in one of the following: 0.2 ml high/low profile 8/12 tube strips (please transfer from 0.5ml PCR tubes if your thermocycler cannot accommodate 0.2 ml tubes) Cut segments of non-skirted 96-well plates Between 31 and 96 Submit samples in one of the following: 0.2 ml high/low profile 8/12 tube strips Cut segments of non-skirted 96-well plates 96-well full-skirted plates (FX robot used for clean-up only accepts full-skirted plates) Please ensure samples are loaded column-wise like in Figure A and not row-wise like in Figure B: A Seq.1 Seq.9 Seq.17 Seq.25 B Seq.2 Seq.10 Seq.18 Seq.26 C Seq.3 Seq.11 Seq.19 Seq.27 D Seq.4 Seq.12 Seq.20 Seq.28 E Seq.5 Seq.13 Seq.21 Seq.29 F Seq.6 Seq.14 Seq.22 Seq.30 G Seq.7 Seq.15 Seq.23 etc. H Seq.8 Seq.16 Seq.24 Figure A. Samples are loaded column-wise. Preferred format

2 A Seq.1 Seq.2 Seq.3 Seq.4 Seq.5 Seq.6 Seq.7 Seq.8 Seq.9 Seq.10 Seq.11 Seq.12 B Seq.13 Seq.14 Seq.15 Seq.16 Seq.17 Seq.18 Seq.19 Seq.20 Seq.21 Seq.22 Seq.23 Seq.24 C Seq.25 Seq.26 Seq.27 Seq.28 Seq.29 Seq.30 D E F G Figure B. Samples are loaded row-wise. (Not ideal because DNA Sequencer customised for columnwise loading.) H If you submit samples that are not in the preferred format, there may be a delay. We assume that samples submitted in 96-well plates are arranged column-wise from left to right (i.e. sample 1 is in well A1, sample 2 in well B1, sample 9 in well A2, etc) If you have deviated from this, please inform us during sample submission. Failure to do so will result in mismatch of sequences to samples. When using 8/12 tube strips, please indicate the first tube of each strip. If you have cut the tubes to use separately, please ensure that when you submit the tubes, the order matches that you have keyed in the CFMS request. Reminder: To avoid mismatch between sequences and samples, please check that the column-wise left to right format is maintained for both physical submission of samples as well as the on-line request on CFMS.

3 Advantages of using 96-well plate/8-tube strips: Only need to label first tube of each strip (1 st, 9 th, 17 th, 25 th ) or first well of plate (assuming samples are organised column-wise from left to right). We will maintain the integrity of the strips or plate and not split the submitted batch during a run. Easy handling for both user and us. Lower risk of sample loss as compared with single 0.2ml tubes Tips in using 8-tube strips: Place 4 empty 0.2 ml tubes at 4 corners of the thermocycler block to reduce pressure from the heated lid otherwise sample tubes may be squashed. Caps for 8-tube strips can be used on 96 well plates.

4 Examples of types of 96-well plates: Please take note that these diagrams were reproduced from the ABgene website. Many other brands are available such as Axygen, USA Scientific, AppliedBiosystems. Some of these are available at the Biopolis Supply Center in B2. AB-0800: Thermo-Fast 96 Skirted For more than 40 samples (can only fit MJ and BioMetra 96-well thermocyclers) AB-0600* Thermo-Fast 96 PCR Plate Non-Skirted For more than 40 samples (for MJ, BioMetra and AppliedBiosystems 96-well thermocyclers). Can be cut to fit 48-well thermocycler. Difference between normal and low-profile 96-well plate:

5 AB-0700*Thermo-Fast 96 PCR Plate Non-Skirted, Low Profile For more than 40 samples(for MJ, BioMetra and AppliedBiosystems 96-well thermocyclers). Can be cut to fit 48-well thermocycler too.

6 SEALING AB-0558 Adhesive PCR Film AB-0626 Adhesive PCR Foil Seal AB-0580 Adhesive Plate Seals AB-3797 Clear Seal 3730

7 AB-3738 Easy Pierce AB-3559 Thermo-Seal AB-0427 Thermo-Mat 96 or AB-0753 Thermo-Mat 384 AB-0770 Thermo-Mat 384 without tabs

8 Caps for 8/12 strips can also be used to seal 96-well plates: AB-0783*Strips of 8 Flat Caps AB-0850*Strips of 12 Flat Caps AB-0265*Strips of 8 Domed Caps AB-0852*Strips of 12 Domed Caps

9 Examples of 8/12 x 0.2ml tubes: AB-0264*Strips of 8 Thermo-Tubes AB-1112 Strips of 12 Thermo-Tubes AB-0783*Strips of 8 Flat Caps AB-0850*Strips of 12 Flat Caps AB-0265*Strips of 8 Domed Caps AB-0852*Strips of 12 Domed Caps

10 Thermocylcers for PCR: Biometra, T1 Thermocycler The MJ Research Tetrad thermocycler MJ THERMAL CYCLER PTC-200

11 MJ Research Dyad thermal Cycler Applied Biosystems GeneAmp PCR 9700 (only half-skirted/non-skirted plates can be used)